Laboratory Methods



GST Fusion Protein Purification from Yeast : This protocol has be used to assay binding of Sst2-RGS and Gbeta/gamma to Gpa1-GST.

TAP Fusion Protein Purification from Yeast

FLAG Fusion Protein Purification from Yeast

Liquid Beta-galactosidase Reporter Gene Assay: This protocol is used to quantitatively measure the pheromone response over a range of pheromone concentrations.

Pheromone Halo Assay : This is a bio-assay that measures the responsiveness of cells to a factor pheromone. The assay is easy to conduct and the results are usually unambiguous and highly reproducible.

SDS Whole Cell Extracts : This is a simple and rapid method for extracting total proteins from Yeast for SDS-PAGE and Western Blotting.

TCA Whole Cell Extracts : This is a quantitative method for extracting total proteins for Western Blotting, especially good with phospho-MAPK antibodies.

Sucrose Density Gradient Fractionation of Yeast Membranes : This is an excellent protocol for localization of membrane associated proteins in yeast.

Beta-galactosidase Filter Assay : This method has been used in our lab to screen for and examine mutants with altered pheromone responses.

"Lazy Bones" PLATE Transformation: This is a quick method for plasmid transformation into yeast.

Storing Yeast Strains

Media Recipies for Yeast

Preparation of Yeast cells for FACS analysis.

Plasmid Isolation from Yeast using Qiagen "QiaPrep" Kit.

Sucrose Density Gradient Fractionation of Yeast Membranes : This is an excellent protocol for localization of membrane associated proteins in yeast.

Plasmid Library Amplification : This protocol has been used successfully in our lab to amplify a LEU2 CEN library. After several screens we determined that the quality of the amplified library was excellent.

Paula's Perfect Immunofluorescence Protocol


Other Protocol Pages
:

The TRAFO high efficiency Yeast Transformation Home Page.

Colony PCR method



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